AU Biomaterials Design Lab:Protocols/Starter Culture Media

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Starter cultures enable us to get a jump-start on culturing bacteria to log-phase while we are expressing proteins. We will usually make 4 starter cultures for our expressions.


  1. Prepare LB in a 250mL Erlenmeyer flask
    1. 0.875g of LB
    2. 35mL of water
  2. Cover the flask with foil
  3. Autoclave
  4. Allow the flask to cool
  5. Add 35uL of 1000X Antibiotic
  6. Inoculate with the bacteria you are culturing using one of the two following methods
    1. Using sterile methods, retrieve a single colony from a petrie dish and drop it into the flask
    2. Using sterile methods, add 5uL of a glycerol stock to the flask
  7. Place the flask in a shaker/incubator and culture overnight at 37C and 200rpm